Environmental Sampling and Assay Device

ABSTRACT

An environmental sampling and assay system, having a coupon storage assembly, storing coupons; an environment sampling assembly, the sampling assembly mixing an environmental sample with a liquid to create a sample-liquid, bearing the environmental sample; a coupon moving assembly; a coupon wetting assembly for automatically wetting a coupon with the sample-liquid; a coupon perceiving device; a data input, adapted to receive a signal; and a data processing and control assembly, controlling the environmental sampling system, the coupon moving assembly, the wetting assembly and the coupon perceiving device. Further, the coupon moving assembly can retrieve a coupon from the coupon storage assembly and move it in a linear manner to the coupon wetting assembly.

RELATED APPLICATIONS

This application is a continuation of application U.S. Ser. No.16/211,744 filed Dec. 6, 2018, which itself is a continuation ofapplication U.S. Ser. No. 15/708,073 filed on Sep. 18, 2017, now U.S.Pat. No. 10,197,558 B1, issued Feb. 5, 2019, and claims benefit ofprovisional application U.S. Ser. No. 62/395,596 filed on Sep. 16, 2016,all of which are incorporated by reference as is fully set forth herein.

BACKGROUND

There is an ongoing concern about the possibility of biological orchemical substances being released into the air with the intent to harmpeople in the release area. Rapid detection of harmful substances isvery helpful in meeting this threat, by speeding evacuation,inoculation, or the administration of an antidote. To meet this threat,with respect to biological substances, the Joint Biological PointDetections System (JBPDS) has been developed. This system includes aninitial sensor that constantly monitors the air, and which triggers anassay test for biological substances, when some warning condition isencountered. Unfortunately, the JBPDS is bulky, and requires cooling andheating, making the entire system even bulkier. Because of thesefactors, it has not been packaged in a form that protects users fromcontamination. Moreover, only one size and shape of assay coupon orstrip can be accepted into the JBPDS, blocking the use of manycommercially available assay strips. Further, the assay reader does notcheck to determine that the assay strip has been properly wetted beforethe time period for an assay-read has elapsed, so that the full assaytime period must elapse before a botched test can be detected. As noted,the JBPDS is configured to only detect biological, as opposed tochemical threats. Accordingly, the use of the JBPDS is hampered by arigid requirement for assay coupons that fit a predetermined geometryand that must be read in a predetermined manner. Personnel using theJBPDS are exposed to the biological substances, for which the JBPDS istesting.

SUMMARY

The following embodiments and aspects thereof are described andillustrated in conjunction with systems, tools and methods which aremeant to be exemplary and illustrative, not limiting in scope. Invarious embodiments, one or more of the above-described problems havebeen reduced or eliminated, while other embodiments are directed toother improvements.

In a first separate aspect, the present invention may take the form ofan environmental sampling and assay system, having a coupon storageassembly, storing coupons; an environment sampling assembly, thesampling assembly mixing an environmental sample with a liquid to createa sample-liquid, bearing the environmental sample; a coupon movingassembly; a coupon wetting assembly for automatically wetting a couponwith the sample-liquid; a coupon perceiving device; a data input,adapted to receive a signal; and a data processing and control assembly,controlling the environmental sampling system, the coupon movingassembly, the wetting assembly and the coupon perceiving device.Further, the coupon moving assembly can retrieve a coupon from thecoupon storage assembly and move it in a linear manner to the couponwetting assembly.

In a second separate aspect, the present invention may take the form ofa system for evaluating wetted coupons, having a camera for takingimages of the coupon and a logic system for evaluating the images.Further, the logic system evaluates the images to determine if thecoupon has been properly wetted.

In a third separate aspect, the present invention may take the form of acoupon wetting system, having a sample cup having a top opening, andholding sample liquid; a pipetting system, including a pipette andreservoir, above the top opening; a coupon storage and movementassembly; and a control system, controlling the pipetting system, andthe coupon storage and movement assembly. Further, wherein the controlsystem controls the pipetting assembly to draw sample liquid through thepipette into the reservoir, and controls the coupon storage and movementassembly to move a coupon to a position directly under the pipette; andcontrols the pipetting system to wet the coupon with the sample fluid.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference to thedrawings and by study of the following detailed descriptions.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1a is a front isometric view of an environmental sampling and assaydevice, according to the present invention.

FIG. 1b is a detail view of FIG. 1, from the same perspective, showingthe wetting assembly.

FIG. 2a is a multichannel biological assay coupon showing the strongpresence of one targeted pathogen.

FIG. 2b is a multichannel biological assay coupon showing the weakpresence of the aforementioned targeted pathogen, or an early indicationof the targeted pathogen.

FIG. 3 is a detail view of FIG. 1, from the same perspective, showingthe coupon storage assembly and coupon moving assembly.

FIG. 4 is a detail view of the device of FIG. 1, from a rotatedperspective, showing further features of the coupon storage assembly.

FIG. 5 is a partial view of the device of FIG. 1 placed into a differentconfiguration in which coupon magazines having differing dimensions holdcoupons having differing dimensions.

FIG. 6 is a block diagram, of the environmental sampling and assaydevice of FIG. 1, showing the communications connections betweendifferent parts of the device.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Definition: In this application the word, “substance” may refer to anorganism, such as a microbe.

For the purpose of promoting an understanding of the principles of theinvention, reference will now be made to the embodiments illustrated inthe drawings and specific language will be used to describe the same. Itwill nevertheless be understood that no limitation of the scope of theinvention is thereby intended. Any alterations and further modificationsin the described embodiments, and any further applications of theprinciples of the invention as described herein are contemplated aswould normally occur to one skilled in the art to which the inventionrelates.

In broad overview, referring to FIGS. 1a, 1b , 3 and 6 in a preferredembodiment of an environmental sampling and assay formation system 10, adata processing and control assembly 12, controls the various elementsof the system 10, to be detailed below, and processes imagery formed bya coupon perceiving device, such as a digital camera 14, to determinepossible presence of biological or chemical contaminants, as will alsobe described further, below. After a detection cycle is triggered byinput received over a data input assembly 68, from a preliminarydetection system 70 (FIG. 6), an environmental sample (air, water orsolid) is mixed with buffer from bottles 16 to form a sample liquidbearing environmental substances held in a sample cup 18 (FIG. 1b ).Bottles 16 also hold deionized water, bleach, and waste, which may alsobe mixed with a sample. A pipetting assembly 20 (which may also bereferred to as a wetting assembly) withdraws sample liquid from thesample cup 18 and delivers sample liquid to coupons 22 (sometimes alsoreferred to as “tickets” in the industry) that are brought from astorage assembly 23, to a wetting position directly below the pipette 24by the coupon movement assembly 26 (FIGS. 3-5). Movement assembly 26then moves the coupons 22 to a position under the camera 14, and couponperceptions, such as digital images, are formed at least every minute,with assembly 12 analyzing these coupon perceptions to detect a couponindication of the presence of a target substance. A human-perceptibleadvisory issuing device issues a signal at any point that such anindication is detected. The advisory may include a visual or auditorycue.

A number of manufacturers produce coupons having various shapes andsizes and are designed to detect various differing biologicalsubstances. Some coupons 22 include an array of detection areas, one foreach of as many as 8 different pathogens, or more. Much of thistechnology is proprietary, so that for many biological substances ofconcern there is only one coupon size (and shape) available that can beused to test for the substance. Accordingly, there is no single couponsize that could be used to detect all biologic substances of concern.Consequently, in order to detect the broadest possible range ofbiological substances, different sized coupons must be accepted.

Referring to FIGS. 5 and 6, to meet this need, storage assembly 23includes differently shaped coupon magazines 30 provided as part ofsystem 10. Each of the magazines 30, however, has a standard shaped base32 (FIG. 3), that is adapted to fit into a standard sized opening 34(shown empty of magazine 30, in FIG. 3) in a support plate 36, therebypermitting a user to exchange first magazine 30, accommodating a stackof first sized coupons 22, for different magazines 30′ and 30″, thataccommodate different sizes of coupon 22′ and 22″. Standard sized base32 and standard sized opening 34 are mating features that permit anysize magazine, having the standard sized base 32 to attach to storageassembly 23 at a standard sized opening 34 (also a mating feature).Other mating features that achieve the same purpose, for examplematching posts and sockets, could serve the same function. Referring toFIG. 3, in like manner, a coupon carriage 40 has standard sized openings42 (FIG. 4), in which can fit a first coupon carrier 44, holding firstsized coupon 22, or a second coupon carrier 44′ adapted to hold thesecond sized coupon 22′, and third sized coupon 22″ (FIG. 5). Althoughmagazines 30 and 30′ and carriers 44 and 44′ appear to be the same inFIG. 3, they may in fact be different, to accommodate the differentmagazines 30, 30′ and 30″ and coupons 22, 22′ and 22″ shown in FIG. 5.Coupons 22 are stacked along their dimension of least extent to permitthe greatest number of coupons 22 to be stored in the magazines 30.

Once system 10 has been configured and is ready to operate, when anindication is received from the preliminary detection system 70, coupons22 that are in a load position at the bottom of the magazine 30 areloaded by linear actuator mechanisms 49, (which are also a part ofstorage assembly 23) from magazines 30 into carriers 44. Gravity causesthe next coupon 22 in each magazine 30 to descend into the load positionfrom which it can be delivered to a carrier 44, next. Coupons 22 arethen moved to a position beneath camera 14 to check for correct couponloading. If this test is passed, pipette assembly 20, which includes anelectronic pipette 24 (controlled by assembly 12), having a disposablereservoir 52 and needle (not shown), extending downwardly from the endof reservoir 52, takes up to 5 cc's of sample liquid from the sample cup18, and uses this to fill the coupon reservoirs for coupons 22. Afterthis filling, the coupons 22 are, for the first few minutes, checked bycamera 14 every 30 seconds to verify proper wetting of the coupon 22,typically by checking to confirm that the control pattern is beginningto appear. If this is not achieved, the test may be aborted, andrestarted, depending on which coupon 22 was not properly wetted and thelogic programming of assembly 12. Alternatively, a human operator isinformed and makes the decision to continue or restart. If the testcontinues (as it generally will) the coupons 22 are placed under camera14 once every minute (illuminated by a light or flash ring 50), therebyproviding enough slack time to fill two sample vials 55 held in carrier56, with as skilled persons will readily recognize, liquid sample.

Many coupons 22 include a control pattern (typically a stripe) thatdevelops when wetted, even in the absence of a target substance, forpurposes of comparison. In a preferred embodiment, this pattern is readby digital camera 14 and used in comparison with the pattern thatdevelops only in the presence of the target substance, in order to forma detection. It is, however, not entirely necessary to compare the testpattern with the control pattern, as in another preferred embodiment, adigitized target pattern (an image of a developed coupon) is introducedinto the memory of assembly 12. This data entry may be performed byplacing a developed coupon 22 or a control section into system 10 duringsystem configuration and using a user interface (not shown) to commandsystem 10 to use a digital camera 14 to take a digital photograph of thedeveloped coupon 22 and store it in memory, properly labeled as adigitized image of a target pattern. In another preferred embodiment,system 10 is provided with digitized target images already stored.Otherwise digitized target images may be introduced into system 10 byway of the data input assembly 68.

Assembly 12 compares each image with the digitized target image storedin its memory, or with the control pattern as perceived by the digitalcamera 14. Although coupon manufacturers specify a development time thatis typically permitted to elapse before a human user reads the coupon,in a preferred embodiment, coupon examination by camera 14 and dataprocessor 12 begins long before this time period has elapsed, with atarget substance detection, also determined minutes before thedevelopment time has passed. In one embodiment, if the target substanceis at a concentration that is at least 20% above the minimum level thatcan be detected by the coupon 22 after the full manufacturer's specifiedcoupon development time has passed, the system provides an advisorysignal prior to the passage of the full manufacturer's specifieddevelopment time. This provides human operators with a quicker resultthat could in some circumstances be very important. In one embodiment,each pixel is compared with a threshold that is one-tenth of theintensity of the fully developed target pattern (dark if the developedtarget pattern is dark and light if the developed target pattern islight) if 95% of the pixels in the target pattern area pass thisthreshold and less than 5% of the pixels outside of the target area passthis threshold, then a detection is determined and a human perceptibleindication, such as an auditory signal and/or visual signal is provided,to alert any nearby people that the target substance has been detected.Many other algorithms, including least squares detection and variouslinear algorithms are used in alternative embodiments. FIG. 2a is anillustration of a test pattern, and FIG. 2b is an illustration of apartially developed coupon 22, showing an indication of the test patternof FIG. 2a . In one preferred embodiment, a coupon test patterndeveloped to the contrast level shown in FIG. 2b is sufficient totrigger an alarm.

The use of digital camera 14 provides a much greater flexibility of use,compared with some prior art systems in which a less robust reader hasbeen used. In a preferred embodiment, assembly 12 is programmed todetect the change in hue that chemical detecting coupons present as anindication of the detection of a chemical substance. Also, carriers 44are provided that can accept the size and shape of chemical coupons.

In one preferred embodiment, system 10 is housed in a vehicle interiorthat is essentially closed to the outside world and with positive airpressure (from air forced in from the outside and thoroughly filtered,on route) causing constant air flow from inside the vehicle interior tothe outside through residual leaks, if any, thereby blocking airbornebiological substances from entering the work area. System 10 is housedin a “glove-box,” a largely transparent, air-tight box, having air-tightgloves sealed to apertures leading through the box walls. Ports leadfrom the glove box to the outside, to permit the gathering of airsamples. Accordingly, a safe workspace is created for users of system10.

The enclosure of the system 10 in an air-tight glove box is not limitedto its use in a vehicle, but is used in many embodiments as it bears theadvantage of protecting test personnel from potential hazards in thesamples, a feature not usually provided in the prior art. One reasonthat prior art systems do not typically afford this level of protectionto test personnel is that an air-tight enclosure may result in thebuildup of water vapor in the glove box due to the handling of waterborne samples. Such handling inevitably leads to evaporation of waterinto the closed glove box volume, creating a risk that watercondensation onto optics or electronics may occur with deleteriouseffects on operation. In a preferred embodiment, the humidity ismonitored by assembly 12 and a dehumidifier is turned on as needed tocreate an optimal or at least not dangerous, humidity level. Due to adesire to maximize operational time and minimize equipment failures, asolid-state dehumidifier using a thermoelectric module and freeconvection heat transfer is preferred, eliminating the need for acompressor or air moving fan. Dehumidifiers of this type are describedat www.myivation.com and are available from Amazon.com under the Ivationtrademark.

In another preferred embodiment, sample cup 18 is sterilized by exposureto ultraviolet light from four LEDs (not shown), which are part of UV-Csterilization system 58. The system 58 is positioned such that both thesample cup 18 and the tops of bottles 16 are sterilized using anintensity of about 96 mW/cm² at 280 nm for 5-20 seconds.

Referring, now, to FIG. 6, system 10 includes the previously noted dataprocessing and control assembly 12, that controls the digital camera 14,the coupon wetting system 20, the coupon storage assembly 23 and thecoupon moving assembly 26 to select a coupon 22, which is then wettedand moved to a position where the digital camera 14, in conjunction withdata processing assembly 12, can monitor its development. Prior to theseactions, however, an environment sampling system 62 must form a liquidsample from ambient air, or from a liquid that is accessible by system10, or even from a solid sample, said liquid sample being then enteredinto a sample cup 18 from which the wetting assembly 20 draws liquid(using a pipette), and uses it to wet a coupon 22. If a target substance(or pathogen) is detected, then an advisory issuing device 64, whichcould be an auditory or visual announcement system, or both, is used tolet an operator know that this has happened. A human input device 66 canbe used by a human operator to command a particular test cycle. In oneembodiment, device 66 is a laptop, tablet or other form of computer thatconnects with the rest of system 10 either through a USB port, ethernetport (which in a preferred embodiment is fiberoptic), or a wirelessconnection such as an RF connection, which may conform to either aBluetooth or WIFI protocol. In another embodiment, device 66 is acustom-made input device, having a keypad and display. In someembodiments, there is overlap between input device 66 and advisoryissuing device 64, with the display screen of device 66 used for theissuance of advisories. In yet another embodiment, as noted above, thepreliminary detection system 70, for example an aerosol particle andbioluminescence detector such as the TacBio trigger developed by the USArmy, is connected to the data input assembly 68 of system 10 and isable to command system 10 to begin a test of a particular type when anaerosol quality is found that meets a set of criteria. For example, whena biological particle count above a specified level is found, or if theparticles have luminescent properties that fit in a prespecified range.The preliminary detection system 70 is fed by an aerosol sampler oraerosol concentrator. In one embodiment, the aerosol concentrator has anair throughput of 4,000 liters per minute and extracts aerosol particlesfrom sampled air and injects them into a secondary circuit flowing at amuch slower rate that is compatible with devices used for creatingliquid samples. Environment sampling system 62 taps into this flowingaerosol concentrate with a 300 liter per minute cyclone wet sampler, toprepare a liquid sample for coupon wetting.

All of the assemblies noted in the discussion above have varyingembodiments not specifically mentioned. In an alternative embodiment,the coupon wetting assembly 20 wets the coupons 22 by way of smalldisposable sponges. The coupon moving assembly 26 makes use of smallelectric vehicles that are optically guided. The coupon storing assembly23 stores coupons 22 on turntables. Moreover, it should be noted thatthere are many forms of digital cameras, including linear cameras thatsimply scan back and forth with one line of pixels, two dimensionaldigital cameras and video cameras, all of which are perceiving devices.In a preferred embodiment, the perceiving device is a fiber optic cable,that is connected to a charge coupled device at the processing andcontrol assembly 12. In a preferred embodiment, processing and controlassembly 12 includes one or more digital computers, which may be in theform of a microcontroller and/or microprocessor, of digital signalprocessing chip or chips. In a preferred embodiment, processing andcontrol assembly 12 includes non-transitory computer readable memorythat has a program that controls the remainder of system 10 to performthe tasks disclosed and claimed herein. Assembly 12, in embodiments,also includes analog-to-digital convertors and digital-to-analogconvertors and amplifiers, sufficient to produce control signals forcontrolling the various systems described.

While a number of exemplary aspects and embodiments have been discussedabove, those possessed of skill in the art will recognize certainmodifications, permutations, additions and sub-combinations thereof. Itis therefore intended that the following appended claims and claimshereafter introduced are interpreted to include all such modifications,permutations, additions and sub-combinations as are within their truespirit and scope.

1. An environmental sampling and assay system, comprising; (a) a couponstorage assembly, storing coupons; (b) an environment sampling assembly,said sampling assembly mixing an environmental sample with a liquid tocreate a sample-liquid, bearing said environmental sample; (c) a couponmoving assembly; (d) a coupon wetting assembly for automatically wettinga coupon with said sample-liquid; (e) a coupon perceiving device; (f) adata input, adapted to receive a signal; (g) a data processing andcontrol assembly, controlling said environmental sampling system, saidcoupon moving assembly, said wetting assembly and said coupon perceivingdevice; and (h) wherein said coupon moving assembly can retrieve acoupon from said coupon storage assembly and move it in a linear mannerto said coupon wetting assembly.
 2. The system of claim 1, wherein saidcoupon storage device stores a first coupon type and a second coupontype, and wherein said first coupon type permits the detection of aspecific pathogen and said second coupon type permits the detection of achemical.
 3. The system of claim 1, wherein control system responds to asaid signal by commanding said coupon storage and movement assembly movea coupon to a location where it can wetted by said wetting system. 4.The system of claim 3, wherein said coupon storage and movement assemblymoves said coupon in a linear manner.
 5. The system of claim 3, whereinsaid coupon storage and movement assembly, includes a coupon storageassembly, and a coupon moving assembly which removes a coupon from saidcoupon storage assembly and moves said coupon to said wetting assembly,when said control system commands said wetting system to wet a coupon.6. The system of claim 1, wherein said coupon storage device stores afirst coupon type and a second coupon type, and wherein said first typeof coupon has a first shape and said second type of coupon has a secondshape.
 7. The system of claim 1, wherein said coupon storage assemblystores said coupon in a vertical magazine, and wherein said couponmoving assembly retrieves a coupon by pushing a bottom most coupon outof said magazine, permitting coupons above it to move down by force ofgravity.
 8. The system of claim 6, wherein said coupon movement assemblypushes said coupon onto a coupon carriage.
 9. The system of claim 7,wherein said coupon carriage also includes a holder for at least onevial.
 10. The system of claim 7, wherein after said coupon movementassembly pushes said coupon onto said coupon carriage said movementassembly moves said coupon to said coupon wetting assembly.
 11. Thesystem of claim 1, wherein said environment sampling system includes aplurality of bottles holding different liquids for mixing with saidenvironmental sample.
 12. A system for evaluating wetted coupons,comprising: (a) a camera for taking images of said coupon; (b) a logicsystem for evaluating said images; and (c) wherein said logic systemevaluates said images to determine if said coupon has been properlywetted.
 13. The system of claim 11, further wherein if there is nodetermination that the coupon has not been properly wetted, said couponis further tested to determine the presence of a substance, but ifimproper wetting is detected said coupon further testing isdiscontinued.
 14. A coupon wetting system, comprising: (a) a sample cuphaving a top opening, and holding sample liquid; (b) a pipetting system,including a pipette and reservoir, above said top opening; (c) a couponstorage and movement assembly; (d) a control system, controlling saidpipetting system, and said coupon storage and movement assembly; and (e)wherein said control system controls said pipetting assembly to drawsample liquid through said pipette into said reservoir, and controls thecoupon storage and movement assembly to move a coupon to a positiondirectly under said pipette; and controls said pipetting system to wetsaid coupon with said sample fluid.
 15. The system of claim 13, furtherwherein after said coupon is wetted said coupon storage and movementassembly moves said coupon away from said pipetting system.
 16. Thesystem of claim 15, further wherein after said coupon is moved away fromsaid pipetting system, said pipetting system fills a sample vial withsaid liquid sample.
 17. The system of claim 15, wherein after saidcoupon is moved away from said pipetting system, said pipetting systemfills two sample vials with said liquid sample.